从猪肠道组织中优化了对隐藏体和介质细胞流体细胞的顺序分离
Hammed Ayansola1, Younggeon Jin2
1Department of Animal and Avian Sciences, University of Maryland.
Journal of visualized experiments : JoVE
|January 6, 2026
概括
这项研究提出了一种新方案,用于从小猪内中分离肠上皮质密室和介质细胞 stromal 细胞 (iMSCs). 这种方法可以同时隔离两种细胞类型,以研究它们的相互作用.
科学领域:
- 胃肠病学 胃肠病学
- 细胞生物学 细胞生物学
- 动物科学动物科学
背景情况:
- 肠表皮和介酶体 stromal 细胞 (iMSCs) 对于肠道的发育和功能至关重要.
- 缺少标准化方法来从猪组织中分离这两种细胞类型.
- 猪模型为人类生理学提供了翻译相关性.
研究的目的:
- 开发和优化一个阶段性协议,从同一个猪骨段逐步分离肠道密室和iMSC.
- 为共同培养研究提供一种可复制的方法,以获得表皮细胞和树皮细胞种群.
- 建立一个研究猪上皮层-层相互作用的基础.
主要方法:
- 使用乙烯基二胺四酸/二二甲醇 (EDTA/DTT) 缓冲器进行密码隔离.
- 采用原酶/丢弃消化来从剩余组织中分离iMSCs.
- 通过细胞培养,免疫染和标记分析 (PDGFRα,CD81) 进行了分离细胞的特征分析.
主要成果:
- 成功地从同一头骨段中分离出可活的肠道密室和iMSC.
- 培养的密室形成了二维 (2D) 体单层.
- 孤立的iMSC粘附在培养瓶上,并表达了关键的 stromal 标记物.
结论:
- 优化的协议有效地从猪内中获得肠上皮和iMSC.
- 这种方法支持使用有机体和共同培养系统对表皮-体相互作用的未来研究.
- 该协议为推进转化研究和动物科学中的猪模型提供了宝贵的工具.
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